Dimerisation of the Yeast K+ Translocation Protein Trk1 Depends on the K+ Concentration
Abstract
:1. Introduction
2. Results
2.1. BiFC Indicates That Trk1[ΔLHL] Can Exist as a Dimer with an Interface Formed by MPMs C and D
2.2. Single Molecule Fluorescence Microscopy and Stepwise Photobleaching
2.3. Quantification of BiFC Fluorescence
3. Discussion
4. Material and Methods
4.1. Strains and Growth Conditions
4.2. Plasmid Construction and Yeast Transformation
4.3. Fluorescence Microscopy
4.4. Single Molecule Fluorescence Microscopy (SMFM)
4.5. Quantification of BiFC
4.6. Molecular Modelling and MD Simulations
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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VC/Trk1[ΔLHL] | VC/Trk1[ΔLHL] | Trk1[ΔLHL]/VC | Trk1[ΔLHL]/VN | ||
---|---|---|---|---|---|
VN/Trk1[ΔLHL] | Trk1[ΔLHL]/VN | VN/Trk1[ΔLHL] | Trk1[ΔLHL]/VC | ||
Result | NO BiFC | BiFC | BiFC | BiFC | Consistent |
Possible arrangement | |||||
Dimer AB/BA | no | no | no | no | no |
Dimer BC/CB | yes | no | no | no | no |
Dimer CD/DC | yes | yes | yes | yes | yes |
Dimer DA/AD | yes | yes | yes | yes | yes |
Tetramer A | yes | no | yes | no | no |
Tetramer B | no | yes | yes | no | no |
Tetramer C | no | yes | yes | yes | no |
Tetramer D | yes | yes | yes | yes | yes |
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Kulik, N.; Kale, D.; Spurna, K.; Shamayeva, K.; Hauser, F.; Milic, S.; Janout, H.; Zayats, V.; Jacak, J.; Ludwig, J. Dimerisation of the Yeast K+ Translocation Protein Trk1 Depends on the K+ Concentration. Int. J. Mol. Sci. 2023, 24, 398. https://doi.org/10.3390/ijms24010398
Kulik N, Kale D, Spurna K, Shamayeva K, Hauser F, Milic S, Janout H, Zayats V, Jacak J, Ludwig J. Dimerisation of the Yeast K+ Translocation Protein Trk1 Depends on the K+ Concentration. International Journal of Molecular Sciences. 2023; 24(1):398. https://doi.org/10.3390/ijms24010398
Chicago/Turabian StyleKulik, Natalia, Deepika Kale, Karin Spurna, Katsiaryna Shamayeva, Fabian Hauser, Sandra Milic, Hannah Janout, Vasilina Zayats, Jaroslaw Jacak, and Jost Ludwig. 2023. "Dimerisation of the Yeast K+ Translocation Protein Trk1 Depends on the K+ Concentration" International Journal of Molecular Sciences 24, no. 1: 398. https://doi.org/10.3390/ijms24010398